Title: Protein Purification/Protein Characterization
Introduction:
Materials & Methods:
Results:
Figure 1. Experimental plate containing PGBR22[G], AV LB Agar, Ampicillin, E. Coli BL21 (DE3), and SOC media with bacterial colonies grown producing purple protein after approximately 27 hours of incubation in a 37 degrees Celsius incubator.
Figure 2. Control plate AV LB Agar, Ampicillin, E. Coli BL21 (DE3), and SOC media without PGBR22 with no visible colonies after approximately 27 hours of incubation in a 37 degrees Celsius incubator.
Figure 3. Fun plate containing AV LB Agar, with swabbed tongue cells from each partner on each half of the petri dish showing visible colonies after approximately 27 hours of incubation in a 37 degrees Celsius incubator.
Figure 4. Two 125 mL Erlenmeyer flasks containing PGEM-gbr22[G], AV LB Agar, Ampicillin, and E. Coli BL21 (DE3) indicating growth of bacteria producing purple protein after 24 hours in a shaking incubator operating at 37 degrees Celsius at 200-350 rpm.
Figure 5. 50 mL conical tube containing purple protein pellet containing transformed bacteria cells (BL21 DE3) with plasmid (pGEM-gbr22[G]). Net weight is 0.54g after centrifuging for 10 minutes at 4 degrees Celsius and 5,000 rpm and removal of supernatant.
Figure 6. 50 mL conical tube containing resuspended purple protein pellet containing transformed bacteria cells (BL21 DE3) with plasmid (pGEM-gbr22[G]), PBS, and lysozyme.
Figure 7. SDS Page Gel of PGEM-gbr22[G] placed on Whatman filter paper covered with cellophane after 1.5 hours of drying at 75 degrees Celsius on the gradient cycle. Wells are seen at the top, with the protein ladder in the first lane, with samples 1-6 from wells 2-7. Purple bands indicate protein found in the sample. The intensity of color represents the amount of the protein in that specific sample.
Figure 8 and 8a. Trial 1 concentration reading of 0.06mg/ml and absorbance reading of 0.013 at 574nm and 0.021 at 280nm for Elution 1.
Figure 9 and 9a. Trial 2 concentration reading of 0.07mg/ml and absorbance reading of 0.014 at 574nm and 0.024 at 280nm for Elution 1.
Figure 10 and 10a. Trial 1 concentration reading of -0.02mg/ml and absorbance reading of 0.005 at 574nm and 0.024 at 280nm for Elution 2.
Figure 11 and 11a. Trial 2 concentration reading of 0.01mg/ml and absorbance reading of 0.006 at 574nm and 0.018 at 280nm for Elution 2.
Introduction:
Materials & Methods:
Results:
Figure 4. Two 125 mL Erlenmeyer flasks containing PGEM-gbr22[G], AV LB Agar, Ampicillin, and E. Coli BL21 (DE3) indicating growth of bacteria producing purple protein after 24 hours in a shaking incubator operating at 37 degrees Celsius at 200-350 rpm.
Figure 6. 50 mL conical tube containing resuspended purple protein pellet containing transformed bacteria cells (BL21 DE3) with plasmid (pGEM-gbr22[G]), PBS, and lysozyme.
Figure 8 and 8a. Trial 1 concentration reading of 0.06mg/ml and absorbance reading of 0.013 at 574nm and 0.021 at 280nm for Elution 1.
Figure 9 and 9a. Trial 2 concentration reading of 0.07mg/ml and absorbance reading of 0.014 at 574nm and 0.024 at 280nm for Elution 1.
Figure 10 and 10a. Trial 1 concentration reading of -0.02mg/ml and absorbance reading of 0.005 at 574nm and 0.024 at 280nm for Elution 2.
Figure 11 and 11a. Trial 2 concentration reading of 0.01mg/ml and absorbance reading of 0.006 at 574nm and 0.018 at 280nm for Elution 2.
Discussion:
Conclusions:
References: